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. 2011 Jul 18;60(8):2041–2050. doi: 10.2337/db11-0042

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© 2011 by the American Diabetes Association.

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details.

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FIG. 5. — Severe diabetes reduces LPL dimers with an increase in Angptl-4. Animals were made moderately or severely diabetic by injecting STZ at a dose of 55 (D55) or 100 mg/kg (D100), respectively, and kept for 4 days. Hearts were isolated and equal amounts of homogenate protein from control (CON), D55, and D100 were loaded onto a heparin–sepharose column and eluted with increasing concentrations of NaCl. LPL activity in the 1.0 mol/L fractions was measured, and peak activity presented as mean ± SE of five animals in each group (A). In a separate experiment, hearts from CON, D55, and D100 were perfused with heparin (5 units/mL), perfusates were collected at the indicated times, and LPL activity was determined (B, representative graph). These perfusates at the indicated times were pooled and loaded onto a heparin–sepharose column. Dimeric LPL in the 1.0 mol/L fractions was determined using TCA precipitation followed by Western blot (B, inset). Results are the mean ± SE of three animals in each group. *Significantly different from control, P < 0.05. Heart homogenates from CON, D55, and D100 were also used for Western blot to detect calnexin (Cn) (C) or Angptl-4 (E) protein expressions normalized by GAPDH. Serum samples were collected from the different groups to detect Angptl-4 using an ELISA assay (D). Angptl-4 mRNA expression from CON and D100 hearts was analyzed by real-time quantitative PCR. Gene expression was evaluated by normalizing to 18S-ribosomal RNA and plotted as fold over control (E, inset). Results are the mean ± SE of five animals in each group. *Significantly different from control, P < 0.05. **Significantly different from control, P < 0.01.